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Title: Uncovering genetic components involved in regulating early immune responses to porcine reproductive and respiratory syndrome (PRRS)

Author
item Lunney, Joan
item Wysocki, Michal
item STEIBEL, JUAN - MICHIGAN STATE
item Kuhar, Daniel
item ERNST, CATHERINE - MICHIGAN STATE
item MCCAW, MONTE - NC STATE UNIVERSITY

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 10/17/2008
Publication Date: 1/10/2009
Citation: Lunney, J.K., Wysocki, M., Steibel, J.P., Kuhar, D.J., Ernst, C.W., Mccaw, M. 2009. Uncovering genetic components involved in regulating early immune responses to porcine reproductive and respiratory syndrome (PRRS). Plant and Animal Genome XVII Conference Abstracts. Available: http://www.intl-pag.org/17/abstracts/P07a_PAGXVII_640.html.

Interpretive Summary: Porcine reproductive and respiratory syndrome virus (PRRSV)-infected pigs are susceptible to pneumonia and reproductive losses. Our goal is to identify the most significant pathways and genes regulating early responses during two pathologic acute PRRSV infections as compared to protective vaccination. For this experiment PRRSV-naïve animals were divided into four groups: (1) pigs infected with acute-type PRRSV MNW2B; (2) pigs infected with acute-type PRRSV NC Powell; (3) pigs vaccinated with a commercial modified-live PRRSV vaccine (Ingelvac ATP®) and (4) control pigs. Tissues [tracheobronchial lymph nodes (TBLN), cranial and caudal lung lobes, and tonsils] were collected between 3-6 days post infection/vaccination. Amplified RNA was labeled with Alexa Fluor® 555 and 647 dyes (Invitrogen) and hybridized to Swine Protein-Annotated Oligonucleotide Microarray www.pigoligoarray.org using a loop design to test for changes in gene expression between all four groups and differences due to day post infection within the PRRSV infected groups. Results obtained for cranial lung tissues affirmed 923, 619 and 747 putatively differentially expressed genes for vaccinated versus control, vaccinated versus infected, and control versus infected, respectively. As expected, pathways involving interferons and other cytokines, as well as chemokines, have been identified as critical for differentiating infected from vaccinated pigs. Final statistical and pathway analyzes are underway to identify the biological functions and regulatory pathways that are most significant. Subsequent experiments will identify candidate genes; RT- PCR will be used to confirm their differential expression. Supported by USDA ARS and CSREES PRRS CAP funds.

Technical Abstract: Porcine reproductive and respiratory syndrome virus (PRRSV)-infected pigs are susceptible to pneumonia and reproductive losses. Our goal is to identify the most significant pathways and genes regulating early responses during two pathologic acute PRRSV infections as compared to protective vaccination. For this experiment PRRSV-naïve animals were divided into four groups: (1) pigs infected with acute-type PRRSV MNW2B; (2) pigs infected with acute-type PRRSV NC Powell; (3) pigs vaccinated with a commercial modified-live PRRSV vaccine (Ingelvac ATP®) and (4) control pigs. Tissues [tracheobronchial lymph nodes (TBLN), cranial and caudal lung lobes, and tonsils] were collected between 3-6 days post infection/vaccination. Amplified RNA was labeled with Alexa Fluor® 555 and 647 dyes (Invitrogen) and hybridized to Swine Protein-Annotated Oligonucleotide Microarray www.pigoligoarray.org using a loop design to test for changes in gene expression between all four groups and differences due to day post infection within the PRRSV infected groups. Results obtained for cranial lung tissues affirmed 923, 619 and 747 putatively differentially expressed genes for vaccinated versus control, vaccinated versus infected, and control versus infected, respectively. As expected, pathways involving interferons and other cytokines, as well as chemokines, have been identified as critical for differentiating infected from vaccinated pigs. Final statistical and pathway analyzes are underway to identify the biological functions and regulatory pathways that are most significant. Subsequent experiments will identify candidate genes; RT- PCR will be used to confirm their differential expression. Supported by USDA ARS and CSREES PRRS CAP funds.